2004 Fiscal Year Final Research Report Summary
Functional analysis of defensins in human innate intestinall immunity.
| Project/Area Number |
15590610
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Asahikawa Medical College |
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Principal Investigator |
AYABE Tokiyoshi Asahikawa Medical College, Department of Gastrointestinal Immunologu and Regenerative Medicine, Associate Professor, 医学部, 寄附講座教員 (90301019)
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| Co-Investigator(Kenkyū-buntansha) |
KOHGO Yutaka Asahikawa Medical College, Third Department of Internal Medicine, Professor and Chairman, 医学部, 教授 (10133183)
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| Project Period (FY) |
2003 – 2004
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| Keywords | innate immunity / antimicrobial peptides / defensins / Paneth cells / host defense / mucosal immunity |
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| Research Abstract |
Small intestinal Paneth cells contribute to mucosal innate immunity by sensing bacteria and secreting microbicidal peptides, particularly alpha-defensins. To investigate the expression and function of Toll-like receptor (TLR)s and related molecules in crypts of human small intestine. Ileal mucosa were obtained from surgical specimens reseated from patients with colon cancer with normal ileum under informed consent. Intact crypts were isolated by EDTA dissociation, and further collagenase dissociation was conducted to prepare crypt single cells. RT-PCR experiments were performed on individual crypts or isolated Paneth cells to assay for alpha-defensins (HD5, HD6), TLR1-10, CD14, MD2 and MyD88 mRNAs. Specific localization was analyzed using anti-HD5 and anti-TLR antibodies in isolated crypts. Isolated intact crypts were exposed to bacteria to induce Paneth cell secretion. Secretions were assayd for bactericidal activity against defensin sensitive S.typhimunum and for HD5 by western blot analysis. Inhibition assay was performed using anti-TLR2 antibody in the secretion assay system. TLR1-8, TLR10, CD14 and MyD88 mRNAs were detected in single isolated crypts and isolated Paneth cells. TLR expression was restricted to Paneth cells in the isolated crypts. Alpha-defensin secretion by Paneth cells in response to bacteria was inhibited by TLR2 antibody pre-treatment. Paneth cells in human small intestinal crypts express Toll-like receptors that appear to be implicated in ex vivo responses to bacterial infection.
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[Book] 自然免疫 vol.12004
Author(s)
綾部時芳
Total Pages
211
Publisher
Innate immunity研究会
Description
「研究成果報告書概要(和文)」より
